Programme, participant list, group lists

for practical sessions, summaries of

presentations and posters

2014-02-18/20, York (GB)

EPPO Workshop on

Accreditation for plant pest

diagnostic laboratories

Table of Contents

Contents of the Brochure

Programme 3

Groups for the Practical Sessions 6

Participant list 7

Abstracts for Oral Presentations 12

Session 1 12

Session 2 15

Session 3 21

Session 4 22

Session 5 24

Pages for notes during practical sessions 26

Abstracts for Posters 29

EPPO Activities on Diagnostics 32

Programme

3

Monday 2014-02-17

Registration 17.30-19.00

Novotel Hotel, Fewster Way, Fishergate, York YO10 4FD

For participants staying in, or near the Novotel, and who will arrive before 19.00 on Monday advance registration is possible at the Novotel. For other participants registration is possible on Tuesday morning at Fera.

Tuesday 2014-02-18

Session 1 General introduction to Quality Assurance and accreditation

08.30-09.00 Registration

09.00-09.15 Fera, GB Welcome and Objectives of the meeting

09.15-9.45 Françoise Petter

EPPO EPPO Standards on Quality Assurance and Interlaboratory Comparisons (Proficiency Testing and Test Performance Studies)

09.45-10.10 Mark

Sykes Proficiency testing group, Fera, GB

Logistics in the Provision of Proficiency Testing

10.10-10.35 David

Galsworthy Fera, GB How the laboratory Quality System can

support and enhance the workings of the plant health diagnostic laboratory

10.35-11.05 Coffee break

Session 2 Experiences of laboratories with establishing accreditation

11.05-11.30 Gunita Bokuma

State Plant Protection Service, LV

Experience with establishing accreditation in the National Phytosanitary Laboratory in Latvia

11.30-11.55 Natalia Sherokolava

All-Russian Plant Quarantine Centre, RU

Russian experience of obtaining accreditation in plant quarantine diagnostics

11.55-12.20 Valerie

Grimault GEVES, FR

Experience of the GEVES laboratory with ISTA since 2001

12.20-12.45 Natasa

Mehle National Institute of Biology, SI

A simple, descriptive assessment of uncertainties of qualitative real time PCR

12.45-13.45 Lunch break

Programme

4

13.45-14.10 Valmir Duarte

Agronomica - Laboratorio de Diagnostico Fitossanitario e Consultoira

Advantages/disadvantages of a private laboratory to meet the quality system: a Brazilian experience

14.10-14-35 Mark

Buimer Naktuinbouw, NL Experience of NAKT with ISO 17025 and ISTA

accreditation

Session 3 Flexible scope in plant pest diagnostic accreditation

14.35-15.00 Marianne

Van Der Blom

National Plant Protection Organization, NL

Phytosanitary focused (flexible) scope

Session 4 Accreditation and new developing diagnostic tools

15.00-15.25 Bart Van De Vossenberg

National Plant Protection Organization, NL

Validation of DNA barcoding tests for the identification of plant pests and plant pathogens in diagnostic laboratories

15.25-15.50 Maja Ravnikar

National Institute of Biology, SI

Determining critical points in validation of isothermal LAMP for accreditation

15.50-16.20 Coffee break

Session 5 Auditing a driver for permanent improvement?

16.20-16.45 Geraldine Anthoine

Laboratoire de la santé des végétaux, FR

Auditing, a driver for continuous improvement of quality management systems?

16.45-17.15 Aude

Chabirand Laboratoire de la santé des végétaux, FR

Statistical analysis in interlaboratory comparison and method validation: Introduction

Poster session – 17.15-18.00

Programme

5

Wednesday and Thursday 2014-02-19/20

Practical sessions and sharing experience sessions

Three parallel practical sessions will be organized. Each session will last half a day as follows:

Wednesday 2014-02-19: 9.00-12.30 and 13.30-17.00

Thursday 2014-02-20: 8.30-12.00

Participants are allocated to different groups (see lists on page 6).

Topic Session Leaders Wednesday

09.00-12.30

Wednesday

13.30-17.00

Thursday

8.30-12.00

Method validation (establishment of a validation plan; example of validation/verification of real-time PCR)

Bart van de Vossenberg, Lynn Laurenson

Group A Group B Group C

Organization of interlaboratory comparisons

Petra Mueller, Sven Inghelbrecht, Natasa Mehle

Group B Group C Group A

Statistical analysis (in interlaboratory comparison and method validation)

Aude Chabirand Geraldine Anthoine

Group C Group A Group B

Thursday 2014-02-20

13.00-16.00

Plenary session with questions and answers on Quality Assurance (including metrology)

Thursday 2014-02-20

16.00-17.00

Conclusions and recommendations

Groups for Practical Sessions

6

Group A

Mr Ahrens Christian

Mr Bryce Steven

Ms Destefanis Marialaura

Mr Douglas Craig

Ms Flint Laura

Mr Hunter Ronald

Ms Jankuviene Lina

Ms Karafla Charikleia

Mr Kornev Konstantin

Ms Lasner Helena

Mr Mernke Dennis

Ms M'rabet Samaali Besma

Mr Nussbaum Ralph-Peter

Mr Oggenfuss Markus

Ms Orosz Szilvia

Ms Ortega Parra Nelia

Mr Riedel Marko

Mr Rigling Daniel

Ms Skomedal Hanne

Ms Telo Patricia

Ms Tomlinson Jenny

Mr Vovkotrub Oleh

Ms Wisdom Rachel

Mr Ziebell Heiko

Group B

Ms Baginska Hanna

Mr Buimer Mark

Mr Choiseul James

Ms Dimova Milena

Ms Dr. Freye-Minks Caroline

Mr Duarte Valmir

Ms Fraser Karen

Ms Gulik Van Der Arina

Ms Harrison Catherine

Ms Kolodziejska Anna

Mr Lacomme Christophe

Ms Leonard Sandrine

Ms Mavrodieva Vessela

Mr Menzel Wulf

Mr Radišek Sebastjan

Ms Schumann Susanne

Ms Sherokolava Natalia

Mr Shneyder Yury

Ms Skelton Anna

Ms Smits-Mastebroek Linda

Ms Stanoevska Mirjana

Ms Wulff Ednar

Group C

Ms Besheva Ani

Ms Bokuma Gunita

Mr Colbert Ruairi

Mr Elphinstone John

Ms Fabich Sabine

Mr Fox Adrian

Mr Galsworthy David

Ms Grimault Valerie

Mr Horky Jaroslav

Mr Jeffries Colin

Mr Nemeth Jozsef

Mr Nowikow Konstantin

Ms Ravnikar Maja

Mr Steyer Stephan

Mr Sykes Mark

Mr Tegel Jukka

Ms Van Der Blom Marianne

Ms Weekes Rebecca

Ms Wilson Veronique

Participant List

7

Participant List

AHRENS Christian (Mr) Institute of Plant Production Sciences, Agroscope Institute of Plant Production Sciences Research Group Molecular Diagnostics, Genomics and Bioinformatics, CH8820 Wädenswil, Switzerland Tel: +41-447836114 - christian.ahrens@agroscope.admin.ch

ANTHOINE Géraldine (Ms) Laboratoire de la santé des végétaux, 7 rue Jean Dixméras, 49044 Angers Cedex 01, France Tel: +33-241207431 - geraldine.anthoine@anses.fr

BAGINSKA Hanna (Ms) Main Inspectorate of Plant Health and Seed Inspection, Central Laboratory, Zwirki i Wigury 73, 87-100 Torun, Poland Tel: +48-566235698 - h.baginska@piorin.gov.pl

BESHEVA Ani (Ms) Bulgarian Food Safety Agency, Central Laboratory for Plant Quarantine, 120 N.Moushanov blvd, 1330 Sofia, Bulgaria Tel: +359-28024707 - a.besheva@bfsa.bg

BOKUMA Gunita (Ms) State Plant Protection Service, Lielvardes str.36/38, LV-1006 Riga, Latvia Tel: +371-67543138 - gunita.bokuma@vaad.gov.lv

BRYCE Steven (Mr) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462603 - steven.bryce@fera.gsi.gov.uk

BUIMER Mark (Mr) Naktuinbouw, sotaweg 22, 2371 gd Roelofarendsveen, Netherlands Tel: +31-713326158 - m.buimer@naktuinbouw.nl

CHABIRAND Aude (Ms) Laboratoire de la santé des végétaux, Ligne Paradis, 7 chemin de l'IRAT, 97410 Saint-Pierrre, La Réunion, Reunion Tel: +262-262382642 - aude.chabirand@anses.fr

CHOISEUL James (Mr) Plant Health Laboratory, Seed Certification Division, Backweston Agri-Labs, Youngs Cross, Celbridge, Co. Kildare, Ireland Tel: +353-16157504 - james.choiseul@agriculture.gov.ie

COLBERT Ruairi (Mr) Department of Agriculture, Food and the Marine, Pesticides, Plant Health and Seed Testing Laboratory Division DAFM, Co. Kildare. IEP Celbridge, Ireland Tel: +353-867712848 - ruairi.colbert@agriculture.gov.ie

DESTEFANIS Marialaura (Ms) Department of Agriculture, Food and the Marine, Backweston campus, Yaoung's cross, 00000 Celbridege, Ireland Tel: +353-16157414 - maria.destefanis@agriculture.gov.ie

DIMOVA Milena (Ms) Central laboratory for plant quarantine, 120 N. Moushanov, 1330 Sofia, Bulgaria Tel: +359-8024776 - mkyuchukova@yahoo.de

DOUGLAS Craig (Mr) Science and Advice for Scottish Agriculture, Roddinglaw Road, Gogarbank, EH12 9FJ Edinburgh, United Kingdom Tel: +44-01312446346 - craig.douglas@sasa.gsi.gov.uk

DR. FREYE-MINKS Caroline (Ms)

LOEWE Biochemica GmbH, Muehlweg 2a, 82054 Sauerlach , Germany Tel: +49-810461620 - caroline.freye@loewe-info.com

DUARTE Valmir (Mr) Agronomica - Laboratorio de Diagnostico Fitossanitario e Consultoira, Rua Ibanez Andre Pitthan Souza, 139, Jardim Itu-Sabara, 91210-070 Porto Alegre, Brazil Tel: +55-5121316262 - valmir@agronomicabr.com.br

Participant List

8

ELPHINSTONE John (Mr) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462334 - john.elphinstone@fera.gsi.gov.uk

FABICH Sabine (Ms) Dienstleistungszentrum Ländlicher Raum Rheinhessen-Nahe-Hunsrück (DLR-RNH), Ruedesheimer Str. 60-68, 55545 Bad Kreuznach, Germany Tel: +49-671820422 - sabine.fabich@dlr.rlp.de

FLINT Laura (Ms) Food and Environment Research Agency, 04GA10, Sand Hutton, York, YO41 1LZ York, United Kingdom Tel: +44-1904462781 - laura.j.flint@fera.gsi.gov.uk

FOX Adrian (Mr) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462747 - adrian.fox@fera.gsi.gov.uk

FRASER Karen (Ms) Science and Advice for Scottish Agriculture, Roddinglaw Road, EH12 9FJ Edinburgh, Scotland, United Kingdom Tel: +44-1312448894 - karen.fraser@sasa.gsi.gov.uk

GALSWORTHY David (Mr) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462261 - david.galsworthy@fera.gsi.gov.uk

GRIMAULT Valerie (Ms) GEVES, 25 rue Georges Morel, CS 90024, 49071 Beaucouze Cedex, France Tel: +33-241225850 - valerie.grimault@geves.fr

GULIK VAN DER Arina (Ms) NAK, Randweg 14, 8304 AS Emmeloord, Netherlands Tel: +31-527635400 - agulik@nak.nl

HARRISON Catherine (Ms) The Food and Environment Research Agency, Sand Hutton York YO41 1LZ, United Kingdom Tel: +44-1904462676 - catherine.harrison@gera.gsi.gov.uk

HORKY Jaroslav (Mr) State Phytosanitary Administration, Division of Diagnostics, Slechtitelu 23, 77900 Olomouc, Czech Republic Tel: +420-585570153 - jaroslav.horky@srs.cz

HUNTER Ronald (Mr) Agrifood & Biosciences Institute Northern Ireland, 18a Newforge Lane Belfast, Northern Ireland BT9 5PX, United Kingdom Tel: +44-02890255253 - ronnie.hunter@afbini.gov.uk

INGHELBRECHT Sven (Mr) ILVO, Plant-Crop Protection, Mycology, Burg.Van Gansberghelaan 96, bus 2, 9820 Merelbeke, Belgium Tel: +32-92722435 - sven.inghelbrecht@ilvo.vlaanderen.be

JANKUVIENE Lina (Ms) The State Plant Service under the Ministry of Agriculture of Republic of Lithuania, Phytosanitary Research Laboratory, Ozo Str. 4A, LT-08200 Vilnius, Lithuania Tel: +370-2616801 - lina.jankuviene@vatzum.lt

JEFFRIES Colin (Mr) Science and Advice for Scottish Agriculture, UK Potato Quarantine Unit, 1 Roddinglaw Road, EH12 9FJ Edinburgh, United Kingdom Tel: +44-1312448868 - colin.jeffries@sasa.gsi.gov.uk

KARAFLA Charikleia (Ms) Benaki Phytopathological Institute, 8 Stefanou Delta Str., 145 61 Athens, Greece Tel: +30-2108180239 - c.karafla@bpi.gr

Participant List

9

KOLODZIEJSKA Anna (Ms) Main Inspectorate of Plant Health and Seed Inspection, Central Laboratory, Zwirki i Wigury Str. 73, 87-100 Torun, Poland Tel: +48-566235698 - a.kolodziejska@piorin.gov.pl

KORNEV Konstantin (Mr) Head of the Bacteriology and molecular methods laboratory of the Federal State Budget Organization, All-Russian Plant Quarantine Center, Pogranichnaya 32, 140150 Bykovo Ramenskoe Region, Russian Federation Tel: +7-9254602907 - konstantin.kornev@gmail.com

LACOMME Christophe (Mr) Science and Advice for Scottish Agriculture, Roddinglaw Road, EH12 9FJ Edinburgh, United Kingdom Tel: +44-1312448916 - christophe.lacomme@sasa.gsi.gov.uk

LASNER Helena (Ms) Agricultural Research Centre, Teaduse 4/6, Saku, 75501 Harju County, Estonia Tel: +372-6729161 - helena.lasner@pmk.agri.ee

LAURENSON Lynn (Ms) Food and Environment Research Agency, Sand Hutton, York, YO41 1DZ, United Kingdom Tel: +44-01904462794 - lynn.laurenson@fera.gsi.gov.uk

LEONARD Sandrine (Ms) Laboratoire Fédéral pour la Sécurité Alimentaire, Chaussée de Namur, 22, 5030 Gembloux, Belgium Tel: +32-498738747 - sandrine.leonard@afsca.be

MAVRODIEVA Vessela (Ms) USDA APHIS Plant Protection and Quarantine, Bldg. 580, BARC-East Powder Mill Rd., 20705 Beltsville, Md, United States Tel: +1-3013139208 - vessela.a.mavrodieva@aphis.usda.gov

MEHLE Natasa (Ms) National Institute of Biology, Vecna pot 111, SI-1000 Ljubljana, Slovenia Tel: +386-59232808 - natasa.mehle@nib.si

MENZEL Wulf (Mr) Leibniz Institute DSMZ, Plant Virus Department Inhoffenstrasse 7B, 38124 Braunschweig, Germany Tel: +49-5312616402 - wum09@dsmz.de

MERNKE Dennis (Mr) Landwirtschaftliches Technologiezentrum Augustenberg, Neßlerstraße 23-31 76227 Karlsruhe, Germany Tel: +49-7219468413 - dennis.mernke@ltz.bwl.de

M'RABET SAMAALI Besma (Ms) General Directorate of Protection and Quality Control of Agricultural Products, Direction Générale de la Protection et du Contrôle de la Qualité des Produits Agricoles Ministère de l'Agriculture 30, rue Alain Savary 1002-Tunis le Belvédère, Tunisia Tel: +216-71788979 - besma.mrabetsamaali@gmail.com

MüLLER Petra (Ms) Julius Kühn Institut (JKI), Federal Research Centre for Cultivated Plants, Institute for National and International Plant Health, Stahnsdorfer Damm 81, 14532 Kleinmachnow, Germany Tel: +49-3320348377 - petra.mueller@jki.bund.de

NEMETH Jozsef (Mr) National Food Chain Safety Office Directorate of Plant Protection, Soil Conservation and Agri-environment, Kodó d. 1., 7634 Pecs, Hungary Tel: +36-72552246 - nemeth.jozsef@baranya.ontsz.hu

NOWIKOW Konstantin (Mr) Justus von Liebig-Labor, Südzucker AG Mannheim/ Ochsenfurt Werk Rain ; Donauwörther Str.50, , 86641 Rain, Germany Tel: +49-9090998215 - konstantin.nowikow@suedzucker.de

Participant List

10

NUSSBAUM Ralph-Peter (Mr) Thüringer Landesanstalt für Landwirtschaft, Referat Pflanzenschutz, Kühnhäuser Straße 101, 99090 Erfurt-Kühnhausen, Germany Tel: +49-36155068124 - ralph-peter.nussbaum@tll.thueringen.de

OGGENFUSS Markus (Mr) Institute of Plant Production Sciences, Agroscope, Research Group Molecular Diagnostics, Genomics and Bioinformatics, CH8820 Wädenswil, Switzerland Tel: +41-447836187 - markus.oggenfuss@agroscope.admin.ch

OROSZ Szilvia (Ms) National Food Chain Safety Office, Budaörsi út 141-145. H-1118 Budapest, Hungary Tel: +36-707013194 - oroszsz@nebih.gov.hu

ORTEGA PARRA Nelia (Ms) Scientia Terrae, Fortsesteenweg 30A Sint-Katelijne-Waver, BE-2860 Sint-Katelijne-Waver , Belgium Tel: +32-15305590 - nop@scientiaterrae.org

RADIšEK Sebastjan (Mr) Slovenian Institute for Hop Research and Brewing, Cesta Zalskega tabora 2, 3310 Žalec, Slovenia Tel: +386-37121626 - sebastjan.radisek@ihps.si

RAVNIKAR Maja (Ms) National Institute of Biology, Vecna pot 111, 1000 Ljubljana, Slovenia Tel: +386-59232801 - maja.ravnikar@nib.si

RIEDEL Marko (Mr) Landesamt für Ländliche Entwicklung, Landwirtschaft und Flurneuordnung , Steinplatz 1, 15806 Zossen, Germany Tel: +49-3370273600 - marko.riedel@lelf.brandenburg.de

RIGLING Daniel (Mr) Swiss Federal Research Institute WSL, Zürcherstrasse 111, 8903 Birmensdorf, Switzerland Tel: +41-447392415 - daniel.rigling@wsl.ch

SCHUMANN Susanne (Ms) Saxon State Company for Environment and Agriculture, Department 65-Phytopathology, Waldheimer Str. 219, House 4, 01683 Nossen, Germany Tel: +49-352426326510 - susanne.schumann@smul.sachsen.de

SHEROKOLAVA Natalia (Ms) "All-Russian Plant Quarantine Centre", Federal State Budget Organization, Pogranichnaya str. 32, 140150 Moscow Oblast, Ramensky Region, Bykovo-2, Russian Federation Tel: +7-4992713824 - natalia_sh@mail.ru

SHNEYDER Yury (Mr) Virology Laboratory, Federal State Budget Organization, All-Russian Plant Quarantine Center, Ramensky region, Bykovo-2, 140150 Moscow Oblast, Russian Federation Tel: +7-9266938580 - yury.shneyder@mail.ru

SKELTON Anna (Ms) Food and environment research agency, 04GA10 Sand Hutton, York, YO41 1LZ, United Kingdom Tel: +44-1904462781 - anna.skelton@fera.gsi.gov.uk

SKOMEDAL Hanne (Ms) the Norwegian Institute for Agricultural and Environmental Research, Høgskoleveien 7, 1430 Ås, Norway Tel: +47-92010068 - hanne.skomedal@bioforsk.no

SMITS-MASTEBROEK Linda (Ms)

NAK, Dutch general inspection service, Randweg 14, 8300 BC Emmeloord, Netherlands Tel: +31-527635400 - lsmits@nak.nl

STANOEVSKA Mirjana (Ms) State Phytosanitary Laboratory, Ministry of Agriculture, Forestri and Water Economy, Str. Aminta III no. 2, 1000 Skopje, Former Yugoslav Republic Of Macedonia Tel: +389-23290638 - mimi_st2007@yahoo.com

Participant List

11

STEYER Stephan (Mr) Centre wallon de Recherches Agronomiques (CRA-W), Wallon Agricultural Research Center, Life Sciences Department, Pest Biology and Biovigilance Unit, Marchal Building, 4 rue de Liroux, BE-5030 Gembloux, Belgium Tel: +32-81620335 - steyer@cra.wallonie.be

SYKES Mark (Mr) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462697 - mark.sykes@fera.gsi.gov.uk

TEGEL Jukka (Mr) Finnish Food Safety Authority Evira, Plant Quarantine Laboratory, Mustialankatu 3, 00790 Helsinki, Finland Tel: +358-405759174 - jukka.tegel@evira.fi

TELO Patricia (Ms) Agronomica - Laboratorio de Diagnostico Fitossanitario e Consultoria, Ibanez Andre Pitthan de Souza, 139, 91210-070 Porto Alegre, Brazil Tel: +55-5121316262 - patricia@agronomicabr.com.br

TOMLINSON Jenny (Ms) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462000 - jenny.tomlinson@fera.gsi.gov.uk

VAN DER BLOM Marianne (Ms) National Plant Protection Organisation, Geertjesweg 15, Postbus 9102, 6700 HC Wageningen, Netherlands Tel: +31-317496852 - m.van.der.blom@minlnv.nl

VAN DE VOSSENBERG Bart (Mr)

Ministry of Economic Affairs, National Plant Protection Organization, National Reference Centre - Molecular Biology, P.O. Box 9102, 6700 HC Wageningen, Netherlands Tel: +31-317496248 - b.t.l.h.van.de.vossenberg@minlnv.nl

VOVKOTRUB Oleh (Mr) State organisation "Central phytosanitary laboratory", 7, Koloskova str., 03138 Kiev, Ukraine Tel: +380-445242702 - vovkotrub@mail.ru

WEEKES Rebecca (Ms) Food and Environment Research Agency, Sand Hutton, YO41 1LZ York, United Kingdom Tel: +44-1904462149 - rebecca.weekes@fera.gsi.gov.uk

WILSON Veronique (Ms) ANSES - Laboratoire de la santé des végétaux, Domaine de Pixérécourt CS 40009, 54220 Malzeville, France Tel: +33-0383290120 - veronique.wilson@anses.fr

WISDOM Rachel (Ms) Plant Health Laboratory, Department of Agriculture, Food and the Marine, Backweston Campus, Celbridge Co.kildare, Ireland Tel: +353-16157363 - rachel.wisdom@agriculture.gov.ie

WULFF Ednar (Ms) Ministry of Food, Agriculture and Fisheries - Danish Veterinary and Food Administration , Soendervang, 4100 Ringsted, Denmark Tel: +45-72276067 - edgaw@fvst.dk

ZIEBELL Heiko (Mr) Julius Kuehn-Institute, Messeweg 11-12, 38104 Braunschweig, Germany Tel: +49-5312993802 - heiko.ziebell@jki.bund.de

MCMULLEN Madeleine (Ms) OEPP/EPPO, 21 boulevard Richard Lenoir, 75011 Paris, France Tel: +33-145207794 - mm@eppo.int

PETTER Françoise (Ms) OEPP/EPPO, 21 boulevard Richard Lenoir, 75011 Paris, France Tel: +33-145207794 - petter@eppo.int

Oral Presentations: Session 1 General Introduction to Quality Assurance and Accreditation

12

Session 1 General introduction to Quality Assurance and accreditation

EPPO Standards on Quality Assurance and Interlaboratory Comparisons (Proficiency Testing and

Test Performance Studies)

Petter Françoise & McMullen Madeleine

European and Mediterranean Plant Protection Organization EPPO, 21 boulevard Richard Lenoir,

75011 Paris, FR, e-mail: petter@eppo.int

In 2010, the Panel on Diagnostics and Quality Assurance suggested that a Standard on

Interlaboratory Comparisons should be developed to provide guidance for laboratories organizing

such activities. A drafting group developed a draft Standard Guidelines for the organization of

interlaboratory comparisons by plant pest diagnostic laboratories. This draft was prepared on the

basis of ISO Standard 17043 Conformity assessment – General requirements for proficiency testing

adapting it to the requirements of plant health laboratories. Some difficulties arose when preparing

guidance on testing for homogeneity and stability of the samples and on statistical analysis. A survey

was organized among the laboratories registered in the EPPO Database on diagnostic expertise and

further progress was made. The draft was sent for country consultation in 2013 and a final version

will be presented for final adoption in 2014.

The EPPO Standard PM 7/98 (1) Specific requirements for laboratories preparing accreditation for a

plant pest diagnostic activity, is one of EPPO’s most often accessed Standards. The revision of this

Standard involved a detailed survey of users from throughout the EPPO region, a survey on widely

used tests (to complete the Appendix), and an Expert Working Group to address the comments.

Input from the Panels on Diagnostics and the collaboration with an expert from the French Forensic

Science laboratory allowed a consensus to be reached on accreditation for morphological

identification. The revised Standard has been sent for Country Consultation.

Oral Presentations: Session 1 General Introduction to Quality Assurance and Accreditation

13

Logistics in the Provision of Proficiency Testing

Sykes Mark

FAPAS, The Food and Environment Research Agency, Sand Hutton, York YO41 1LZ, UK,

mark.sykes@fera.gsi.gov.uk

FAPAS is an accredited and internationally renowned proficiency testing provider, with over 23

years’ experience. The focus of FAPAS is on food chemistry but FAPAS also runs schemes for food

microbiology, genetically modified materials, and drinking water chemistry and microbiology. FAPAS

has also run some plant health proficiency tests (PT) and non-food chemistry tests. There are many

considerations for running a PT scheme. Technically, these include test material production,

homogeneity and verification testing, packaging and stability testing, dispatch of test samples,

statistical analysis of PT results, setting acceptance criteria, and defining assessment types. Equally

important considerations include providing clear instructions to participants, managing participant

address and contact details, maintaining confidentiality, choice of courier, capturing results and

method information, managing data handling, responding to enquiries, and use of expert advice. All

this has to be considered even before applying for accreditation for the scheme. This presentation is

intended to provide food for thought for the workshop discussions. Examples will be used to

illustrate some of the problems that FAPAS has encountered and the ways in which these have been

overcome.

Oral Presentations: Session 1 General Introduction to Quality Assurance and Accreditation

14

How the laboratory Quality System can support and enhance the workings of the plant health

diagnostic laboratory

Galsworthy, David

Head of Quality at the Food and Environment Research Agency (FERA), GB

e-mail: David.Galsworthy@fera.gsi.gov.uk

This introductory talk will cover ideas on how the quality system of the laboratory can be designed

and operated in ways that support the laboratory as well as complying with the ISO 17025 quality

Standard. Practical examples of how this can be achieved will be presented drawing upon the

experience gained whilst carrying out 1500 assessment visits for the UK accreditation body as well as

the design and operation of the FERA quality system. Examples will include smart ways for document

control, use of process mapping tools and the application of a flexible scope of accreditation.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

15

Session 2 Experiences of laboratories with establishing accreditation

Experience with establishing accreditation in the National Phytosanitary Laboratory in Latvia

Bokuma Gunita State Plant Protection Service, National Phytosanitary Laboratory, Lielvardes street 36/38, Riga, LV-1006, Latvia email: gunita.bokuma@vaad.gov.lv

In 2001 a decision was made to implement a quality system in the National Phytosanitary Laboratory in Latvia and the process of preparation for accreditation took place from 2001 to 2003. Another important decision had to be made when it came to the choice of methods/pests to be proposed for accreditation. There were many methods for testing a wide range of pests and diseases in the laboratory and priorities for crops and pests for which our results would have the most significant impact had to be chosen. After a time and labour-consuming preparation process in 2003 all the necessary documents were submitted to the Latvian National Accreditation Bureau. After elimination of all non-conformities, accreditation for six methods was received in 2005. The quality management system was implemented and applies to the whole laboratory. Advantages of implementation of the quality system were firstly that the demands of State Plant Protection Service management and a client of the laboratory Sanitary Border Inspection of Latvia were fulfilled. Other advantages are order in the laboratory, ordered documents, traceability of samples, clearly determined responsibilities, awareness concerning tests performed and results obtained.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

16

Russian experience of obtaining accreditation in plant quarantine diagnostics

Sherokolava Natalia All-Russian Plant Quarantine Center 32 Pogranichnaya str., Bykovo, Moscow oblast, Russia email: natalia_sh@mail.ru

Two years ago, a reform of the accreditation system took place in the Russian Federation. The purpose was to harmonize activities in the field of technical regulations with the world trends. It was a very important step towards international recognition of the performance of accredited laboratories in Russia. As a result of this reform a single government body for accreditation was created. Quarantine phytosanitary diagnostics is a specific area of laboratories’ activity. This causes difficulties in the process of accreditation both for laboratories and for the accreditation body. All these problems can be solved only by joint efforts.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

17

Experience of GEVES laboratory with ISTA accreditation since 2001.

Grimault Valerie

GEVES, 25 rue Georges Morel, CS 90024, 49071 Beaucouzé – France

email: Valerie.GRIMAULT@geves.fr

GEVES laboratory has been ISTA accredited since 2001. The ISTA accreditation system will be

presented and compared to the ISO 17025 and ISO 9001 accreditation systems which GEVES also

has. Examples of the implications of these accreditation systems in the laboratory will be presented

in terms of the quality system, traceability, validation of the ability to perform a method,

participation and organization of proficiency tests and audits.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

18

A simple, descriptive assessment of uncertainties of qualitative real time PCR

Nataša Mehle (1), Tanja Dreo (1), Maja Ravnikar (1) 1) National Institute of Biology, Večna pot 111, SI-1000 Ljubljana, Slovenia email: natasa.mehle@nib.si

In most cases tests used for plant pest diagnosis give qualitative results despite being based on measurement of some kind e.g. fluorescence level in real-time PCR. While this result is just one part of the diagnostic process it may be critical in its influence on the final result if it is subject to high uncertainty. To be able to identify critical points in a diagnostic protocol a laboratory should attempt to assess the uncertainty of each step and its relative importance in the whole process. A relatively simple process for estimating level of uncertainty for a diagnostic test based on qualitative real-time PCR will be presented. An example of real-time PCR testing of grapevine yellows phytoplasmas will be used to illustrate generation of data in a comprehensive and self-explanatory table, summarizing possible impacts on the final results and measures for reducing the uncertainty. The approach identifies critical points in the whole process (from sampling to the final reports), and allows the estimation of the relative importance of each step. This approach has been used in support of accreditation according to the ISO 17025 in our laboratory since 2012 and can be applied to other detection methods and pests.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

19

Advantages/disadvantages of a private laboratory to meet the quality system: a Brazilian experience

Duarte Valmir; Telo, Patricia de Souza Agronomica, Rua Ibanez Andre Pitthan Souza, 139, Jardim Itu-Sabara, Porto Alegre, RS, Brazil, 91210-070 email: valmir@agronomicabr.com.br

Agronomica is a private laboratory authorized, based on NBR ISO/IEC 17025, by the Ministry of Agriculture (MAPA), working on phytosanitary diagnostics since May 2006 in Brazil. Its scope includes bacteria, fungi, phytoplasmas, viruses, viroids, nematodes, mites, insects, molluscs and weeds. Located in Southern Brazil, it receives samples from all over the country. In 2013, 17 000 samples were analyzed, including all sorts of plant material, import, export, and internal controls. Its main customer is MAPA. The laboratory team consists of 13 people with plant health expertise, four consultants, and 11 support staff. The main advantage is the flexibility in the management of all activities, facilitating the targeting and strengthening (such as hiring staff, buying new equipment and identification kits, and or personal training) of any area to meet new demands. The disadvantage is the cost to keep everything under a quality management system, to maintain a qualified staff, to obtain permits to import type strains of quarantine pests etc.. New rules for laboratory accreditation and monitoring by MAPA (Normative Instruction # 57, Dec 11, 2013) were established. Agronomica’s challenge in 2014 will be to start the accreditation process by accrediting a few tests in the National Metrology Institute.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

20

Experience of Naktuinbouw laboratories with ISO 17025 and ISTA accreditation

Buimer Mark

Naktuinbouw laboratory, Sotaweg 22, 2371 GD Roelofarendsveen,NL,

e-mail: m.buimer@naktuinbouw.nl

The experience of Naktuinbouw laboratories with ISO 17025 and ISTA accreditation is presented in

this talk. Stichting Nederlandse Algemene Kwaliteitsdienst Tuinbouw (the Netherlands Inspection

Service for Horticulture) is generally known as Naktuinbouw. Naktuinbouw promotes and monitors

the quality of produce, processes and chains in horticulture. The main focus is on propagating

material, of national and international origin. Naktuinbouw is an Autonomous Public Authority (APA)

regulated by the Ministry of Economic Affairs. In the obligatory inspection system, Naktuinbouw

applies the prescribed European directives and legislation for propagating material for floricultural,

arboricultural and vegetable crops. These directives are anchored in Dutch legislation in the form of

the Netherlands Seeds and Planting Materials Act. Naktuinbouw is an independent and unbiased

organization. Public duties relating to basic inspections that are the responsibility of other quality

and/or inspection services (national and international) are not performed by Naktuinbouw or only

performed on a cooperative basis. Naktuinbouw Laboratories support these public duties and

operate according to strict protocols and quality assurance systems, the ISO 17025 and ISTA

accreditation. New tests and protocols, when needed, are developed by our own Research &

Development department.

Oral Presentations: Session 2 Experiences of laboratories with establishing accreditation

21

Session 3 Flexible scope in plant pest diagnostic accreditation

Phytosanitary focused (flexible) scope

Van der Blom, Marianne

National Plant Protection Organisation, Catharijnesingel 59, 3511 GG Utrecht, NL E-mail: m.van.der.blom@minlnv.nl

The National Reference Centre of the Dutch NPPO (NRC) is ISO/IEC 17025 accredited for a fixed

scope for five specific organism/host plant combinations. However, regarding the wish to extend the

accreditation, both fixed and standard flexible scopes do not fit well with the nature of the work. The

focus of the NRC is on diagnoses for thousands of different organism/host plant combinations and

not on routine testing. Some diagnoses are made only once every few years. Full validations and

specific internal control programmes for all individual tests as required under traditional scopes are

therefore not feasible. To fit all diagnostic activities under accreditation, the NRC is investigating the

possibilities for a special phytosanitary flexible scope. This is done in consultation with the Dutch

Accreditation Board. In such ‘phyto-flex scope’, the diagnostic process is considered the basis.

Verification of tests and use of quality controls combined with assurance of the diagnostic process

and competence of specialists have to warrant the quality of the diagnoses.

Oral Presentations: Session 4 Accreditation and new developing diagnostic tools

22

Session 4 Accreditation and new developing diagnostic tools

Validation of DNA barcoding tests for the identification of plant pests and plant pathogens in diagnostic laboratories

van de Vossenberg Bart(1) & Westenberg Marcel(1) & Botermans Marleen(1) 1) Dutch National Plant Protection Organization (NPPO-NL), P.O. Box 9102, 6700 HC Wageningen, The Netherlands, e-mail: b.t.l.h.van.de.vossenberg@minlnv.nl

DNA barcoding is a diagnostic method that uses short standardised genetic markers in an organism’s DNA to aid identification of regulated plant pests and pathogens. In contrast to specific conventional and real-time PCR tests, it does not provide a qualitative black and white result but will allow for generic identification on the required taxonomical level. For this reason, DNA barcoding is increasingly used in plant health laboratories. As is the case for any diagnostic test, DNA barcoding tests have to be validated and verified when used under an ISO 17025 quality assurance system. Validation is the confirmation that a test is fit for purpose by examination and by providing objective evidence. The majority of steps in a DNA barcoding protocol (i.e. DNA extraction, PCR and Sanger Sequencing) can be validated following guidelines in EPPO Standard PM 7/98 Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity. Generating consensus sequences is an important part of DNA barcoding and depends heavily on the expertise of technicians handling raw data. This step is monitored using 2nd and 3rd line control samples rather than being validated. For the sequence analysis step, a single validation is not sufficient as databases with constantly changing content are used for identification. Sequence analyses are therefore "validated" each time an identification is performed.

Oral Presentations: Session 4 Accreditation and new developing diagnostic tools

23

Determining critical points in validation of isothermal LAMP for accreditation

Maja Ravnikar(1), R. Lenarčič(1), D. Morisset(1), Nataša Mehle(1), Ion Gutierrez (1) Polona

Kogovšek(1), Manca Pirc (1) and Dreo Tanja (1)

1) National Institute of Biology, Vecna pot 111, SI-1000 Ljubljana, Slovenia

email: maja.ravnikar@nib.si; tanja.dreo@nib.si

Increased global trade with fresh plant material offers additional possibilities for plant pathogens to spread around the world efficiently. Early detection of pathogens in infected material at the entry points (such as ports, airports, border crossings) and production sites can prevent spread of pests and possible consequent economic loss. Early detection is particularly important in the case of easily transmissible pathogens like Potato spindle tuber viroid (PSTVd) and Ralstonia solanacearum (Rs). Here we describe accurate detection of Rs and PSTVd using LAMP isothermal amplification that gives fast results including simple sample preparation. The test can be performed in a laboratory using qPCR or using a touch screen operated portable device that detects amplification in real-time based on fluorescence, suitable for onsite detection. The use of intercalating dye enables an additional confirmation: melting curve analysis of the amplicon. LAMP tests are approximately ten times less sensitive than real-time PCR and ten times more sensitive than conventional RT-PCR and are less prone to inhibitors than PCR methods. Critical points in LAMP validation will be presented and discussed.

Oral Presentations: Session 5 Auditing a driver for permanent improvement?

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Session 5 Auditing a driver for permanent improvement?

Auditing, a driver for continuous improvement of quality management systems?

Anthoine Geraldine

ANSES - Plant Health Laboratory, 7 rue Jean Dixmeras, 49044 Angers, FR,

geraldine.anthoine@anses.fr

Auditing consists of verifying the conformity of processes or operations to rules or procedures

described in the laboratory and in specific standards. Auditing is one of the activities required by ISO

17025 (§ 4.14), as an internal supervision of the quality management system. However, auditing is

also an external activity, either on the demand of the laboratory or as an obligation for accreditation.

Guidelines and requirements for auditing are described in ISO 19011. Auditing is generally based on

the collection of proof and elements of traceability which are compared to internal or external

requirements. The results of this comparison lead to improvements of the quality system being

identified, e.g. by modifying current procedures or developing new ones. The quality of

communication is one of the key elements for the success of auditing. The different types of audits

and their contribution to the continuous improvement of the quality system will be presented.

Practical examples will be given. The limit of audits in the long term will also be discussed.

Oral Presentations: Session 5 Auditing a driver for permanent improvement?

25

Statistical analysis in interlaboratory comparison and method validation: Introduction

Chabirand Aude (1), Folcher Laurent (2), Hérau Vincent (3)

1) Anses Plant Health Laboratory, 7 chemin de l’IRAT, 97410 Saint-Pierre, Reunion Island, FR,

aude.chabirand@anses.fr

2) Anses Plant Health Laboratory, Domaine de la Motte au Vicomte, 35653 Le Rheu, FR,

laurent.folcher@anses.fr

3) Anses Plant Health Laboratory, 7 rue Jean Dixméras, 49044 Angers, FR, vincent.herau@anses.fr

Interlaboratory comparisons and method validations in plant pathology require the use of statistics.

This presentation is an introduction to the practical session “Statistics analysis in interlaboratory

comparison and method validation”. It presents the basic concepts in statistics for their correct

application in plant pathology. A statistical unit is a unit of observation or measurement for which

data are collected. In statistics, a sample is a group of statistical units (usually individuals) selected

from a larger group (the population). By studying the sample (descriptive statistics), it is hoped to

draw valid conclusions about the population (inferential statistics). This generalization, which is

possible under certain conditions, is an example of the application of statistical modelling. It is

essential to identify the nature of the variables studied from the sample to determine the

appropriate data processing and apply the adequate statistical tests. The presentation focuses on

the different types of variables, the main probability distributions, the sampling and the different

statistical tests for proportion or mean comparisons. All these concepts are presented and applied in

the context of plant pathology.

Notes for Practical Sessions

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Posters

29

POSTERS

Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight

Bühlmann Andreas (1), Pothier Joël F. (1,2), Smits Theo H.M. (1,2), Andreou Michael (3), Oggenfuss Markus (1), Ahrens Christian H. (1), Boonham Neil (4), Duffy Brion (1,2) and Frey Jürg E. (1) 1) Agroscope, Institute of Plant Production Sciences, Molecular Diagnostics, Genomics and Bioinformatics, CH8820 Wädnswil, Switzerland; 2) Zurich University of Applied Sciences, Environmental Genomics and Systems Biology Group, CH8820 Wädenswil, Switzerland; 3) OptiGene, Horsham, West Sussex, RH13 5QR, United Kingdom; 4) The Food and Environment Research Agency FERA, Sand Hutton, YO41 1LZ York, United Kingdom

The aim of this study was to demonstrate the utility of a novel diagnostic target identification strategy based on comparative genomics and bioinformatics. The plant pathogenic bacterium Erwinia amylovora, which causes fire blight on trees of the subfamily Spiraeoideae was chosen as a test pathosystem. 14 genomes of E. amylovora were used to define a core Erwinia genome using the software EDGAR. The 2974 core genes were then screened using BLAST against 10 genomes of related bacteria and against the entire NCBI nucleotide database to identify true singleton genes, i.e. genes specific for Erwinia. Several potential target genes identified by this pipeline were validated with Loop-mediated isothermal amplification (LAMP), a novel isothermal amplification method with increased sensitivity and speed. The assay for EAMY3195 showed the best sensitivity and speed and was validated on more than 100 artificially inoculated greenhouse samples and on more than 100 naturally infected field samples. Besides its specificity the most important advantage of LAMP is its speed, which allows positive detection of a bacterial sample in as little as 15 minutes. LAMP assays thus have significant potential as fast pest detection assays for insects, bacteria and other pest organisms at point of entries such as airports.

Posters

30

EPPO database on diagnostic expertise

Petter, Françoise & McMullen Madeleine

EPPO, 21 Bld Richard Lenoir, Paris 75011, FR.

In 2004, the EPPO Council stressed that the implementation of phytosanitary regulations for quarantine pests was being jeopardized by decreasing expertise in plant protection and declared a state of emergency for Plant Health. The Panel on Diagnostics proposed that an inventory should be made of the available expertise on diagnostics in Europe. In order to address some of these concerns, a database on the diagnostic expertise available in the EPPO region was launched in 2006.

The database includes an inventory of the diagnostic expertise available in the EPPO region on: Pests of EPPO A1 and A2 Lists; Pests mentioned in EPPO Standards PM4: Production of Healthy Plants for Planting; Pests possibly presenting a risk to EPPO member countries (EPPO Alert List); Plants on the EPPO List of invasive alien plants. The database does not include expertise on common pests which are widely distributed in the EPPO region.

The section ‘laboratory list’ includes: Laboratories sorted by country; Administrative details; List of experts in the laboratory; Information on quality programmes, accreditation, possibility of offering training. The section ‘expertise list’ includes expertise sorted by: level of expertise: species, genus (expert who can identify most species in a genus), family (expert who can identify most species in a family)…; names of organisms in alphabetical order (simply choose a letter); types of methods used by the experts (morphological, serological, molecular, bioassay, biochemical, fatty acids or classical microbiology).

Validation data is also included for diagnostic tests for regulated pests. The validation data (generated by various laboratories in EPPO member countries) are presented according to a common format. Validation data can be submitted by any laboratory registered in the EPPO database on diagnostic expertise. To be added to the database a laboratory must be part of the NPPO diagnostic network. Laboratories can register in the database by contacting diagnostics@eppo.int.

Posters

31

Management and automation of qPCR diagnostic workflow according to QA standards

M. Hren(2), K. Gruden(1), K. Zupančič(2), N. Mehle(1), U. Čepin(2) and M. Ravnikar(1) 1) National Institute of Biology, Večna pot 111, 1000 Ljubljana, Slovenia 2) BioSistemika Ltd., Tehnološki park 24, 1000 Ljubljana, Slovenia E-mail: maja.ravnikar@nib.si

A growing challenge in laboratories dealing with high throughput real-time PCR (qPCR) analyses for diagnostic purposes is how to make the complete process from sample preparation to data analysis and interpretation faster without compromising traceability. This poster presents a case study on detection of bacteria with qPCR in complex plant samples, focusing on automation, unification, traceability and simplification of the complete process. The new procedure combines an automated simple and quick homogenization step and an automated DNA extraction method based upon the binding of DNA to magnetic beads. Further dry-lab steps (experiment design, creation of templates for lab work, data analysis, results interpretation and reporting) that are complex, repetitive, time consuming and typically involve use of different software and paper documents were connected into an innovative software solution GENEIO®, an advanced Electronic Laboratory Notebook, developed by BioSistemika LLC. This is an easy-to-operate workflow environment installed in a centralised server and accessible to all employees with different levels of permissions via a safe browser connection from anywhere inside the local network. It communicates with LIMS and qPCR thermal cyclers and stores all the necessary data in one place (quality assurance compliant) including raw Cq and fluorescence data, QC-related data, time stamps etc. In this way an overview and control over all qPCR work inside the organizational unit (laboratory, department, organization etc.) is allowed for the laboratory managers. Manual wet-lab steps – laborious loading of mastermixes and samples onto qPCR plates have been further shortened and controlled using a smart pipetting assistant developed by BioSistemika LLC. (PlatR TM). Combining such modern approaches enables control over the whole workflow, save huge amounts of time without compromising traceability and relieves key personnel, which is of extreme importance for high throughput routine qPCR analyses.

EPPO Activities on Diagnostics

32

EPPO activities on diagnostics

http://www.eppo.int/QUARANTINE/diagnostic_activities.htm

EPPO Activities on Diagnostics

33